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A Missense in HSF2BP Causing Primary Ovarian Insufficiency Affects

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A Missense in HSF2BP Causing Primary Ovarian Insufficiency Affects RESEARCH ARTICLE A missense in HSF2BP causing primary ovarian insufficiency affects meiotic recombination by its novel interactor C19ORF57/BRME1 Natalia Felipe-Medina1†, Sandrine Caburet2,3†, Fernando Sa´ nchez-Sa´ ez1, Yazmine B Condezo1, Dirk G de Rooij4, Laura Go´ mez-H1, Rodrigo Garcia-Valiente1, Anne Laure Todeschini2,3, Paloma Duque1, Manuel Adolfo Sa´ nchez-Martin5,6, Stavit A Shalev7,8, Elena Llano1,9, Reiner A Veitia2,3,10*, Alberto M Penda´ s1* 1Molecular Mechanisms Program, Centro de Investigacio´n del Ca´ncer and Instituto de Biologı´a Molecular y Celular del Ca´ncer (CSIC-Universidad de Salamanca), Salamanca, Spain; 2Universitede Paris, Paris Cedex, France; 3Institut Jacques Monod, Universitede Paris, Paris, France; 4Reproductive Biology Group, Division of Developmental Biology, Department of Biology, Faculty of Science, Utrecht University, Utrecht, Netherlands; 5Transgenic Facility, Nucleus platform, Universidad de Salamanca, Salamanca, Spain; 6Departamento de Medicina, Universidad de Salamanca, Salamanca, Spain; 7The Genetic Institute, "Emek" Medical Center, Afula, Israel; 8Bruce and Ruth Rappaport Faculty of Medicine, Technion, Haifa, Israel; 9Departamento de Fisiologı´a y Farmacologı´a, Universidad de Salamanca, Salamanca, Spain; 10Universite´ Paris-Saclay, Institut de Biologie F. Jacob, Commissariat a` l’Energie Atomique, Fontenay aux Roses, France *For correspondence: [email protected] (RAV); [email protected] (AMP) Abstract Primary Ovarian Insufficiency (POI) is a major cause of infertility, but its etiology remains poorly understood. Using whole-exome sequencing in a family with three cases of POI, we †These authors contributed equally to this work identified the candidate missense variant S167L in HSF2BP, an essential meiotic gene. Functional analysis of the HSF2BP-S167L variant in mouse showed that it behaves as a hypomorphic allele Competing interests: The compared to a new loss-of-function (knock-out) mouse model. Hsf2bpS167L/S167L females show authors declare that no reduced fertility with smaller litter sizes. To obtain mechanistic insights, we identified C19ORF57/ competing interests exist. BRME1 as a strong interactor and stabilizer of HSF2BP and showed that the BRME1/HSF2BP Funding: See page 26 protein complex co-immunoprecipitates with BRCA2, RAD51, RPA and PALB2. Meiocytes bearing Received: 17 March 2020 the HSF2BP-S167L variant showed a strongly decreased staining of both HSF2BP and BRME1 at Accepted: 26 August 2020 the recombination nodules and a reduced number of the foci formed by the recombinases RAD51/ Published: 26 August 2020 DMC1, thus leading to a lower frequency of crossovers. Our results provide insights into the molecular mechanism of HSF2BP-S167L in human ovarian insufficiency and sub(in)fertility. Reviewing editor: Bernard de Massy, CNRS UM, France Copyright Felipe-Medina et al. This article is distributed under Introduction the terms of the Creative Commons Attribution License, The process of gametogenesis is one of the most complex and highly regulated differentiation pro- which permits unrestricted use grams. It involves a unique reductional cell division, known as meiosis, to generate highly specialized and redistribution provided that cells: the gametes. Indeed, the outcome of meiosis is the production of oocytes and spermatozoa, the original author and source are which are the most distinctive cells of an adult organism and are essential for the faithful transmis- credited. sion of the genome across generations. Felipe-Medina et al. eLife 2020;9:e56996. DOI: https://doi.org/10.7554/eLife.56996 1 of 39 Research article Cell Biology The meiotic division is an orderly process that results in the pairing and synapsis of homologous chromosomes and crossover (CO) formation, which ultimately enable homologous chromosomes segregation (Hunter, 2015; Loidl, 2016; Zickler and Kleckner, 2015). In mammals, pairing of homologs is dependent on the repair of self-induced double-strand breaks (DSBs) during prophase I by homologous recombination (Handel and Schimenti, 2010) and it leads to the intimate alignment of homologous chromosomes (synapsis) through the zipper-like synaptonemal complex (SC) (Cahoon and Hawley, 2016). The SC is a proteinaceous tripartite structure that provides the struc- tural framework for DSBs repair (Baudat et al., 2013), as epitomized by the tight association of the recombination nodules (RNs, multicomponent recombinogenic factories) and the axial elements of the SC (Zickler and Kleckner, 2015). Meiotic DSBs repair is an evolutionarily conserved pathway that is highly regulated to promote the formation of at least one CO per bivalent. This chromosome connection between bivalents through chiasmata is required for a correct reductional division. As other DNA repair processes, proper meiotic recombination is essential for genome stability and alterations can result in infertility, miscarriage and birth defects (Geisinger and Benavente, 2017; Handel and Schimenti, 2010; Webster and Schuh, 2017). Infertility refers to failure of a couple to reproduce and affects 10–15% of couples (Isaksson and Tiitinen, 2004). Infertility can be due to female factors, male factors, a combination of both or to unknown causes, each category representing approximately 25% of cases (Isaksson and Tiitinen, 2004; Matzuk and Lamb, 2008). There are several underlying causes and physiological, genetic and even environmental and social factors can play a role. Forward and reverse genetic analyses in model organisms have identified multiple molecular pathways that regulate fertility and have allowed to infer reasonable estimates of the number of protein-coding genes essential for fertility (de Rooij and de Boer, 2003; Schimenti and Handel, 2018). Primary ovarian insufficiency (POI) is a major cause of female infertility and affects about 1–3% of women under 40 years of age. It is characterized by cessation of ovarian function before the age of 40 years. POI results from a depletion of the ovarian follicle pool and can be isolated or syndromic. Genetic causes of POI account for approximately 20% of cases (Rossetti et al., 2017). Although infertility-causing pathogenic variants are inherently unlikely to spread in a population, they can be observed within families, especially when there is consanguinity. Such cases provide crucial insights into the function of the genes and molecular mechanisms that they disrupt. Over the last decade, causative variants in several genes have been found using whole exome sequencing in ‘POI pedi- grees’. In particular, pathogenic variants in genes involved in DNA replication, recombination or repair, such as STAG3, SYCE1, HFM1, MSH5 and MEIOB have been formally implicated in this condi- tion by ourselves and others (Caburet et al., 2014; Caburet et al., 2019a; de Vries et al., 2014; Guo et al., 2017; Primary Ovarian Insufficiency Collaboration et al., 2014). In this study, we have identified in a consanguineous family with POI the candidate S167L mis- sense variant in HSF2BP, an essential yet poorly studied meiotic gene. HSF2BP encodes an interactor of the heat-shock response transcription factor HSF2 (Yoshima et al., 1998). During the course of this work and, in agreement with our results, two independent groups showed that HSF2BP is essen- tial for meiotic recombination through its ability to interact with BRCA2 (Brandsma et al., 2019; Zhang et al., 2019). Here, we report that the introduction of the missense variant HSF2BP-S167L in mouse leads to subfertility and DNA repair defects during prophase I. In addition, we identified a protein complex composed of BRCA2, HSF2BP, and the as yet unexplored C19ORF57/BRME1 (mei- otic double-stranded break BRCA2/HSF2BP complex associated protein) as a key component of the meiotic recombination machinery. Our studies show that a single substitution (S167L) in HSF2BP leads to a reduced loading of both BRME1 and HSF2BP at the RNs. Furthermore, our results suggest that meiotic progression requires a critical threshold level of HSF2BP/BRME1 for the ulterior loading of the recombinases to the RNs. Results Clinical cases The parents are first-degree cousins of Israeli Arab origin. Of the five daughters, three are affected with POI and presented with early secondary amenorrhea. They had menarche at normal age (at 13– Felipe-Medina et al. eLife 2020;9:e56996. DOI: https://doi.org/10.7554/eLife.56996 2 of 39 Research article Cell Biology 14) but with irregular menses that stopped around 25. Only one of the patients affected by POI could have a child with the help of a fertility treatment (see pedigree in Figure 1). In order to identify the genetic basis of this familial POI case, we performed whole exome sequencing on genomic DNA from two POI patients, III-2 and III-3, and their fertile sister III-10 (Supplementary file 1a). Variants were filtered on the basis of (i) their homozygosity in the patients, (ii) their heterozygosity or absence in the fertile sister, (iii) their absence in unrelated fertile in-house controls and (iv) a minor allele fre- quency (MAF) below 0.01 in all available databases (Supplementary file 1b). This filtering process led to the identification of a missense substitution located in the HSF2BP gene: rs200655253 (21:43630396 G > A, GRCh38). The variant lies within the sixth exon of the reference transcript ENST00000291560.7 (NM_007031.2:c.500C > T) and changes a TCG codon into a TTG (NP_008962.1:p.Ser167Leu). It is very rare (Variant Allele Frequency/VAF 0.0001845 in the GnomAD database and 0.0005 in the GME
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